In 1999, California Air Resources Board (CARB) implemented a regulation that required all gasoline cars sold in California be fitted with an Onboard Refueling Vapor Recovery System (ORVR). The ORVR system is designed to prevent Volatile Organic Compounds (VOCs) from escaping into the atmosphere during refuelling by storing the gas vapours in a carbon canister. Due to the complex nature of the fuel system, making design changes could have large implications on the ORVR performance of the vehicle. It is therefore desirable to develop a CFD model that can predict the effects of design changes, thereby reducing the need to perform physical tests on each design iteration. This master thesis project was performed at the Fuel Systems department at Volvo Cars in order to help reduce project lead times and product development costs by incorporating CFD as a part of the fuel system development cycle. The CFD results obtained were validated through experimental tests that were also performed as part of this project. In this master thesis project, a CFD model was developed to simulate the refuelling of gasoline for a California specification Volvo XC90 with an OPW-11B pump pistol. The model was set up in STAR-CCM+ using the Eulerian Volume of Fluid model for multiphase flow, the RANS realizable k-epsilon turbulence model and the two layer all y+ wall treatment. The effects of the carbon canister were modelled as a porous baffle interface in the simulations where viscous and inertial resistances of the porous media were adjusted to obtain a desired pressure drop across the canister. This method proved to be a suitable simplification for this study. The effects of evaporation as well as a chemical adsorption model for the carbon canister have been excluded from the project due to time limitations. It was found that the CFD simulations were in good agreement with the experimental results, especially with respect to capturing the overall behaviour of the fuel system during refuelling. It was found that resolving the flow spatially (and temporally) in the filler pipe was a crucial part in ensuring solver stability. A pressure difference between experiment and simulation was also observed as a consequence of excluding evaporation from the CFD model. After the CFD model had been verified and validated, changes to different parts of the fuel system were investigated to observe their effects on ORVR performance. These included changing the recirculation line diameter, changing the carbon canister properties and changing the angle of how the pump pistol was inserted into the capless unit. It was found that the recirculation line diameter is a very sensitive design parameter and increasing the diameter would result in fuel vapour leaking back out into the atmosphere. Similarly, increasing the back pressure by swapping to a different carbon canister would result in the leakage of fuel vapour. On the other hand, insignificant changes in system behaviour were observed when the fuel pistol angle was changed.

In 1999, California Air Resources Board (CARB) implemented a regulation that required all gasoline cars sold in California be fitted with an Onboard Refueling Vapor Recovery System (ORVR). The ORVR system is designed to prevent Volatile Organic Compounds (VOCs) from escaping into the atmosphere during refuelling by storing the gas vapours in a carbon canister. Due to the complex nature of the fuel system, making design changes could have large implications on the ORVR performance of the vehicle. It is therefore desirable to develop a CFD model that can predict the effects of design changes, thereby reducing the need to perform physical tests on each design iteration. This master thesis project was performed at the Fuel Systems department at Volvo Cars in order to help reduce project lead times and product development costs by incorporating CFD as a part of the fuel system development cycle. The CFD results obtained were validated through experimental tests that were also performed as part of this project. In this master thesis project, a CFD model was developed to simulate the refuelling of gasoline for a California specification Volvo XC90 with an OPW-11B pump pistol. The model was set up in STAR-CCM+ using the Eulerian Volume of Fluid model for multiphase flow, the RANS realizable k − ε turbulence model and the two layer all y + wall treatment. The effects of the carbon canister were modelled as a porous baffle interface in the simulations where viscous and inertial resistances of the porous media were adjusted to obtain a desired pressure drop across the canister. This method proved to be a suitable simplification for this study. The effects of evaporation as well as a chemical adsorption model for the carbon canister have been excluded from the project due to time limitations. It was found that the CFD simulations were in good agreement with the experimental results, especially with respect to capturing the overall behaviour of the fuel system during refuelling. It was found that resolving the flow spatially (and temporally) in the filler pipe was a crucial part in ensuring solver stability. A pressure difference between experiment and simulation was also observed as a consequence of excluding evaporation from the CFD model. After the CFD model had been verified and validated, changes to different parts of the fuel system were investigated to observe their effects on ORVR performance. These included changing the recirculation line diameter, changing the carbon canister properties and changing the angle of how the pump pistol was inserted into the capless unit. It was found that the recirculation line diameter is a very sensitive design parameter and increasing the diameter would result in fuel vapour leaking back out into the atmosphere. Similarly, increasing the back pressure by swapping to a different carbon canister would result in the leakage of fuel vapour. On the other hand, insignificant changes in system behaviour were observed when the fuel pistol angle was changed.In 1999, California Air Resources Board (CARB) implemented a regulation that required all gasoline cars sold in California be fitted with an Onboard Refueling Vapor Recovery System (ORVR). The ORVR system is designed to prevent Volatile Organic Compounds (VOCs) from escaping into the atmosphere during refuelling by storing the gas vapours in a carbon canister. Due to the complex nature of the fuel system, making design changes could have large implications on the ORVR performance of the vehicle. It is therefore desirable to develop a CFD model that can predict the effects of design changes, thereby reducing the need to perform physical tests on each design iteration. This master thesis project was performed at the Fuel Systems department at Volvo Cars in order to help reduce project lead times and product development costs by incorporating CFD as a part of the fuel system development cycle. The CFD results obtained were validated through experimental tests that were also performed as part of this project. In this master thesis project, a CFD model was developed to simulate the refuelling of gasoline for a California specification Volvo XC90 with an OPW-11B pump pistol. The model was set up in STAR-CCM+ using the Eulerian Volume of Fluid model for multiphase flow, the RANS realizable k − ε turbulence model and the two layer all y + wall treatment. The effects of the carbon canister were modelled as a porous baffle interface in the simulations where viscous and inertial resistances of the porous media were adjusted to obtain a desired pressure drop across the canister. This method proved to be a suitable simplification for this study. The effects of evaporation as well as a chemical adsorption model for the carbon canister have been excluded from the project due to time limitations. It was found that the CFD simulations were in good agreement with the experimental results, especially with respect to capturing the overall behaviour of the fuel system during refuelling. It was found that resolving the flow spatially (and temporally) in the filler pipe was a crucial part in ensuring solver stability. A pressure difference between experiment and simulation was also observed as a consequence of excluding evaporation from the CFD model. After the CFD model had been verified and validated, changes to different parts of the fuel system were investigated to observe their effects on ORVR performance. These included changing the recirculation line diameter, changing the carbon canister properties and changing the angle of how the pump pistol was inserted into the capless unit. It was found that the recirculation line diameter is a very sensitive design parameter and increasing the diameter would result in fuel vapour leaking back out into the atmosphere. Similarly, increasing the back pressure by swapping to a different carbon canister would result in the leakage of fuel vapour. On the other hand, insignificant changes in system behaviour were observed when the fuel pistol angle was changed.

LORVR ou obstruction récurrente des voies respiratoires (ORVR) est la cause la plus fréquente de maladie pulmonaire chronique chez le cheval adulte. La maladie se caractérise par une hyperréactivité bronchique, une production excessive de mucus et une inflammation neutrophilique pulmonaire qui ont pour effet de réduire la compliance dynamique du poumon et daugmenter la résistance des voies respiratoires au débit aérien. Alors que la maladie est une entité documentée dans la littérature depuis plus de 200 ans, les mécanismes moléculaires qui la sous-tendent restent incomplètement élucidés à ce jour. De précédentes études ont montré l'implication de variations d'expression génique, pour la plupart grâce à l'utilisation combinée de la transcription inverse et de la réaction en chaîne par polymérase (RT-PCR). Cette technique bien connue ne permet l'étude que d'un nombre limité de gènes en simultané. Les microdamiers apparaissent comme la technique de choix pour l'étude de l'expression génique à large échelle. A lheure où cette étude a débuté, il nexistait pas de microdamier dexpression spécifiquement dédié au cheval sur le marché. Un microdamier hétérologue humain (Human Genome U133 Plus 2.0 GeneChip, Affymetrix®) a donc été utilisé pour tenter didentifier de nouveaux gènes dintérêt pour comprendre la pathogénie de lORVR chez le cheval. L'utilisation du microdamier a permis de détecter 46 gènes dont lexpression variait d'un facteur supérieur à 4 entre chevaux sains et atteints. Parmi les gènes mis en avant par le microdamier, trois gènes (CYBB, MARCKS et PTX3) présentaient un intérêt particulier au regard de la littérature et leur variation dexpression a été confirmée par RT-PCR quantitative (RT-qPCR). La pentraxine 3 (PTX3) notamment appartient à la composante humorale de la réponse immune innée. Elle agit comme un ancêtre fonctionnel des anticorps. En effet, elle reconnaît les microbes, active le complément et facilite la reconnaissance de certains pathogènes spécifiques, dont Aspergillus fumigatus (A. fumigatus), par les phagocytes. La suite du travail sest donc focalisée sur PTX3. LADN complémentaire (ADNc) codant pour PTX3 et lADN codant pour le second exon de PTX3 ont été clonés et séquencés. LADN codant pour le second exon de PTX3 équin présentait une délétion de 120 paires de bases par rapport à la séquence humaine. Par ailleurs, un épissage alternatif du second exon de PTX3 a été mis en évidence, résultant en deux formes de la protéine, "épissée" (32 kilodaltons) et "complète" (42 kilodaltons). Afin d'étudier les effets de lexposition à des poussières de foin sur l'expression de PTX3 dans les voies respiratoires, six chevaux sains et six chevaux atteints dORVR ont été soumis pendant dix jours à un challenge environnemental aux poussières de foin. Les chevaux ont été étudiés via des tests cliniques, fonctionnels et biochimiques. Les lavages broncho-alvéolaires (LBA) ont été récoltés lors de l'initiation du protocole et après les dix jours d'exposition aux poussières de foin. Par ailleurs, des sections bronchiques ont été réalisées chez deux chevaux atteints d'ORVR en crise et euthanasiés pour raisons éthiques et deux chevaux indemnes d'affections respiratoires provenant de l'abattoir. PTX3 était détectée dans les macrophages du LBA après exposition aux poussières de foin et dans les cellules épithéliales bronchiques. PTX3 était surexprimée dans les macrophages du LBA chez tous les chevaux après dix jours d'exposition aux poussières de foin. Cependant, le niveau de PTX3 était plus élevé dans le surnageant des cellules provenant des chevaux atteints d'ORVR que dans celui provenant des cellules des chevaux sains. Par ailleurs, il semble que PTX3 soit surexprimée dans les cellules épithéliales bronchiques chez le cheval atteint d'ORVR. Cette piste est intéressante sur le plan thérapeutique puisque l'activation de l'expression de PTX3 dans les cellules épithéliales bronchiques est sous le contrôle du facteur c-Jun N-terminal kinase (JNK), un facteur différent de celui qui contrôle l'expression de PTX3 dans les leucocytes, à savoir le facteur nucléaire kappa B (NF-κB). L'effet de différents stimuli sur la production et lexcrétion de PTX3 par les cellules du LBA a été étudié ex-vivo après mise en culture et stimulation à la poussière de foin, au lipopolysaccharide (LPS), à A. fumigatus et avec une solution saline tamponnée pour le phosphate (PBS), utilisée comme contrôle. La présence de PTX3 a été mesurée par Western Blot six heures et vingt-quatre heures après les différentes stimulations dans les cellules et dans les surnageants respectivement. PTX3 était détectée dans les cellules et dans les surnageants mais aucune différence significative d'expression na pu être mise en évidence que ce soit entre les différents stimuli, entre les différents temps ou avec le contrôle. Ces résultats suggèrent que la poussière de foin est un inducteur indirect de PTX3./ Recurrent airway obstruction (RAO) is the first respiratory disease in adult horses. Environmental causes of RAO are well described, but the molecular mechanisms of the disease remain unclear. Previous studies have highlighted the implications of variations in gene expression in the disease, most using reverse transcription polymerase chain reaction (RT-PCR). This well-known technique limits the number of genes that can be studied in a single assay. The microarray appears to be the platform of choice for massively parallel gene expression profiling and provides a good tool for exploratory research. However, at the time of the experiment, there was no equine-specific microarray available on the market. Because they are commercially available, highly specific and well annotated, human and mouse large-scale microarrays are an exploratory alternative to equine-specific microarrays. The first part of the present study was performed to highlight new targets not previously related to the disease and able to improve our understanding of the molecular mechanisms of the disease. A human microarray (Human Genome U133 Plus 2.0 GeneChip, Affymetrix®) was used to study gene expression in nucleated cells originating from peripheral blood and bronchoalveolar lavage fluid (BALF) in RAO-affected horses. A total of 46 candidates were identified with differentially regulated genes between RAO-affected horses and controls. Based on their documented function, five of these genes were selected for the real-time quantitative RT-PCR (RT-qPCR) validation procedure: CYBB, MARCKS and PTX3. The RT-qPCR results confirmed those obtained with the microarray, pointing out these genes as new directions for future experiments. In particular, the long pentraxin 3 (PTX3) plays an important role in the host defence and over-expression of PTX3 may contribute to airways injury. Therefore, it was hypothesised that PTX3 may have relevance in the understanding of the pathogenesis of RAO. In the second part of the study, PTX3 and its implication in RAO in horses were characterized in more details. Six healthy horses and six RAO-affected horses were submitted to a dusty environment challenge. PTX3 DNA and cDNA were cloned and sequenced. A 120bp deletion was found in the DNA of the second exon of equine PTX3 in comparison to human PTX3. In addition, an alternative splicing of the second exon of PTX3 occurred, resulting in two forms of the protein: "spliced" (32 kilodaltons) and "full length" (42 kilodaltons). The pentraxin domain was very well conserved in the two forms with 94% of amino-acids conserved between equines and humans. The effects of dust exposure and subsequent RAO crisis on PTX3 expression in the BALF and the bronchial epithelial cells were studied. Dust exposure induced PTX3 in vivo in BALF macrophages both in healthy horses and in RAO-affected horses. However, PTX3 was over-expressed in the BALF supernatant and, interestingly, in the bronchial epithelial cells from RAO-affected horses in crisis. Considering epithelial cells as one of the major cell types in the airways, PTX3 produced at this level may play an important role in the disease process. The effects of different components (ie lipopolysaccharide and A. fumigatus) and different concentrations of hay dust suspension upon PTX3 expression and excretion in BALF cells have been tested ex-vivo and compared to control (ie. Phosphate-Buffered Saline). PTX3 was detected in cells and cell culture supernatant under all conditions but there was no difference between conditions including the control. These results suggest that dust is an indirect inducer of PTX3.